Morpholinylpyridone compounds

ABSTRACT

The invention provides novel morpholinylpyridone compounds of formula (I), pharmaceutical compositions containing such compounds, and methods for using such compounds in treatment of diseases including cancer, type II diabetes, inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders and viral infections; 
     
       
         
         
             
             
         
       
     
     wherein R 1 , R 2  and A are as defined in the specification.

CROSS REFERENCE

This application is a continuation application of U.S. patent application Ser. No. 16/639,902, filed on Feb. 18, 2020, which is a § 371 national phase of International Application No. PCT/EP2018/072791, filed on Aug. 23, 2018, which claims priority and benefit of European Patent Application No. 17187567.7, filed on Aug. 23, 2017, each of which are incorporated by reference herein.

FIELD OF THE INVENTION

The invention provides novel morpholinylpyridone compounds of formula (I), pharmaceutical compositions containing such compounds, and methods for using such compounds in treatment of diseases including cancer and type II diabetes.

BACKGROUND OF THE INVENTION

Enzymes belonging to the family of phosphatidylinositide 3-kinases (PI3K) are regulators of several important cellular events. The family consists of three classes, 1, 11 and Ill and while the Class I group has been an interesting drug target for many years, Class II and Ill are less exploited. The PI3K Class III, vacuolar protein sorting 34 (Vps34, PIK3C3) forms a heterodimer with its regulatory subunit p150 (Vps15) and this dimer takes part in several complexes regulating vesicular trafficking events such as autophagy, endocytosis, exocytosis and micropinocytosis (Amaravadi et al. Clin Cancer Res. 2011, 17:654-666; Carpentier et al. 2013, Traffic). The enzyme is responsible for phosphorylation of phosphatidylinositol (PI) to phosphatidylinositol (3)-phosphate (PI3P). The ligand binding to PX and FYVE domains results in recruiting and delocalization of these effector proteins that lead to vesicular formation, elongation and movement (Backer et al. J Biochem. 2008, 410:1-17).

Autophagy is a catabolic process where cellular components are targeted for degradation by enclosing them in double-membrane vesicles, auto-phagosomes that are fused with the protease-containing lysosomes. This is a mean for the cell to handle damaged organelles and misfolded proteins and by that maintain cellular function. The pathway is also a way of recirculating cellular content into new building blocks (Boya et al, Nat Cell Biol 2013, 15; 713-720). Autophagy is a cellular response to stressful conditions as nutrient deprivation, acidosis and hypoxia but also to drug treatment. Therefore, autophagy inhibition is a means to potentiate cancer drugs and resensitize drug resistant tumors (Nagelkerke et al, Semin Cancer Biol 2014, 31; 99-105). Most advanced tumors show a high upregulation of autophagic flux (Leone et al. Trends in Endocrin Metab 2013, 24; 209-217). An established marker for studying autophagic flux is the detection of autophagic puncta in the form of lipidated LC3 protein on the autophagosome. Inhibition of Vps34 results in the inhibition of autophagy as measured by LC3 redistribution into puncta (Dowdle et al., Nat Cell Biol 2014, 16; 1069-79).

As recently described, ablation of the regulatory subunit p150 leads to increased insulin sensitivity in vivo due to decreased insulin receptor internalization (Nemazanyy, Nature Commun., 2015, 6:8283). A kinase dead heterozygous animal model confirms this result with increased glucose tolerance and increased insulin sensitivity (WO2013076501).

Several disease states could benefit from Vps34 inhibition including cancer, inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders, type II diabetes and viral infections (Reviewed in Rubinsztein et al, Nat Rev 2012, 11; 709-730). Cancer forms that would benefit from Vps34 inhibition include, but are not limited to, breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, renal cancer, colon cancer, glioma, prostate cancer, ovarian cancer, melanoma, and lung cancer as well as hypoxic tumors. There is thus a need for novel and potent inhibitors of Vps34.

Previous disclosures describing Vps34 inhibitors in use to affect diseases include WO2015150555; WO2015150557; WO2015108861; WO2015108881; WO2012085815; WO2012085244; WO2013190510; Farkas, J. Biol. Chem., 2011 286(45) 38904-12.

DESCRIPTION OF THE INVENTION

An object of the invention is to provide novel and potent inhibitors of Vps34. Another object of the invention is to provide novel and potent inhibitors of Vps34 that may be used for treating cancer and other diseases such as type II diabetes.

According to one aspect of the invention, there is provided a compound of formula (I)

wherein

-   R¹ is C₁-C₃alkyl or cyclopropyl; -   R² is selected from hydrogen, C₁-C₃haloalkyl and C₁-C₃alkyl; -   A is

wherein

-   R³ is selected from R⁶, C₁-C₆alkyl, amino, N—C₁-C₃alkylamino,     N,N-diC₁-C₃alkylamino and C₁-C₃alkoxyC₁-C₃alkyl, said C₁-C₆alkyl and     said C₁-C₃alkoxyC₁-C₃alkyl being optionally substituted with one R⁶     and/or one or more halo; -   R⁴ is selected from C₁-C₆alkyl, C₁-C₆alkoxy, C₁-C₆haloalkyl,     C₃-C₆cycloalkyl and phenyl, said phenyl being optionally and     independently substituted with one of more of fluoro, chloro,     methyl, methoxy, dimethylamino, trifluoromethoxy, trifluoromethyl,     cyclopropyl; -   R⁵ is selected from halogen, C₁-C₆alkyl, C₁-C₆alkoxy, C₁-C₆haloalkyl     and C₃-C₆cycloalkyl; -   R⁶ is selected from phenyl, monocyclic heteroaryl, C₃-C₆cycloalkyl,     heterocyclyl, each optionally substituted with one or more R⁷; -   R⁷ is selected from halogen, amino, N—C₁-C₃alkylamino,     N,N-diC₁-C₃alkylamino and C₁-C₃alkoxyC₁-C₃alkyl, C₁-C₃alkoxy,     C₁-C₃haloalkoxy, C₃-C₆cycloalkyl, C₁-C₃haloalkyl and C₁-C₃alkyl;     or a pharmaceutically acceptable salt thereof.

According to one embodiment of this aspect of the invention, R² is selected from hydrogen and C₁-C₃alkyl, such as hydrogen.

According to one embodiment of this aspect of the invention, R¹ is methyl.

According to one embodiment of this aspect of the invention, R⁷ is selected from fluoro, cyclopropyl and methyl.

According to one embodiment of this aspect of the invention, R⁷ is fluoro or methyl.

According to one embodiment of this aspect of the invention, R⁴ is selected from methyl, trifluoromethyl, cyclopropyl and phenyl, said phenyl being optionally meta-substituted with one of fluoro, chloro, methyl, methoxy, dimethylamino, trifluoromethoxy, trifluoromethyl and cyclopropyl; and R⁵ is selected from chloro, cyclopropyl, methyl and trifluoromethyl.

According to one embodiment of this aspect of the invention, R⁴ and R⁵ are independently selected from C₁-C₃haloalkyl, such as C₁-C₃fluorooalkyl, such as trifluoromethyl.

According to one embodiment of this aspect of the invention, R⁴ is selected from methyl, trifluoromethyl and cyclopropyl; and

R⁵ is selected from chloro, cyclopropyl, methyl and trifluoromethyl.

According to one embodiment of this aspect of the invention, R³ is selected from R⁶, C₁-C₃alkyl, N,N-diC₁-C₃alkylamino and methoxyC₁-C₃alkyl, said C₁-C₃alkyl being optionally substituted with one R⁶.

According to one embodiment of this aspect of the invention, R⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, isoxazolyl, pyrazolyl, oxazolyl, cyclopropyl, cyclopentyl, pyrrolidinyl and tetrahydrofuryl, each optionally substituted with one or more R⁷.

According to one embodiment of this aspect of the invention, R⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, pyrazolyl, cyclopropyl, pyrrolidinyl, piperidinyl, and tetrahydrofuryl, each optionally substituted with one or more R⁷

According to one embodiment of this aspect of the invention, R⁶ is selected from phenyl, pyridyl, pyrrolidinyl, pyrazolyl, tetrahydrofuryl, each optionally substituted with one or more R⁷.

According to one embodiment of this aspect of the invention, R⁶ is selected from

According to one embodiment of this aspect of the invention, R⁶ is selected from

According to one embodiment of this aspect of the invention. R⁶ is selected from

According to one embodiment of this aspect of the invention, R³ is selected from

According to one embodiment of this aspect of the invention, R³ is selected from

According to one embodiment of this aspect of the invention, R³ is selected from

According to one embodiment of this aspect of the invention, A is

According to one embodiment of this aspect of the invention, A is

According to one embodiment of this aspect of the invention, R¹ is methyl or cyclopropyl;

-   R² is hydrogen; -   A is

and R³ is selected from

According to one embodiment of this aspect of the invention, R¹ is methyl;

-   R² is hydrogen; -   R⁴ and R⁵ are CF₃; -   A is

and R³ is selected from

According to one embodiment of this aspect of the invention, R¹ is methyl;

-   R² is hydrogen; -   R⁴ is CF₃; -   A is

and R³ is selected from

According to one embodiment of this aspect of the invention,

-   R¹ is methyl or cyclopropyl; -   R² is hydrogen; -   R⁴ and R⁵ are CF₃; -   A is

and R³ is selected from

According to one embodiment of this aspect of the invention, said compound is 4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

-   6-[4-[(4-Fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   6-[4-[(5-Fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide; -   6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   6-[4-Cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   6-[4-(1,2-Dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   6-[4-(1-methylcyclopropyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-1H-pyridin-2-one; -   N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)benzenesulfonamide;     or a pharmaceutically acceptable salt thereof.

According to one embodiment of this aspect of the invention, said compound is

-   4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   6-[4-[(4-Fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   6-[4-[(5-Fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; -   N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide; -   6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; -   4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;     or pharmaceutically acceptable salt thereof.

In one aspect of the invention, there is provided a compound according to the present invention, for use in the treatment or prophylaxis of a disease.

In one aspect of the invention, there is provided a compound according to the present invention, for use in treating cancer. Typically, said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, renal cancer, colon cancer, glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors.

In one aspect of the invention, there is provided a compound according to the present invention, for use in treating type II diabetes.

In one aspect of the invention, there is provided a compound according to the present invention, for use in treating a disease selected from inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders and viral infections.

In one aspect of the invention, there is provided use of a compound according to the present invention, in the preparation of a medicament for treating cancer. Typically said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, renal cancer, colon cancer, glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors.

In one aspect of the invention, there is provided use of a compound according to the present invention, in the preparation of a medicament for treating type II diabetes.

In one aspect of the invention, there is provided use of a compound according to the present invention, in the preparation of a medicament for treating a disease selected from inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders and viral infections.

In one aspect of the invention, there is provided a method of treating cancer, comprising administering a therapeutically effective amount of a compound according to the present invention, to a patient in need thereof. Typically, said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, renal cancer, colon cancer, glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors.

In one aspect of the invention, there is provided a compound according to the present invention, for use in treating cancer, wherein said cancer treatment further comprises radiation therapy.

In one aspect of the invention, there is provided a method of treating cancer, comprising administering a therapeutically effective amount of a compound according to the present invention, to a patient in need thereof, in conjunction with radiation therapy.

The compounds of the present invention may also be employed in cancer treatment in conjunction with radiation therapy and/or surgical intervention.

Generally, the use of cytotoxic and/or cytostatic agents in combination with a compound or composition of the present invention will serve to:

-   (1) yield better efficacy in reducing the growth of a tumor or even     eliminate the tumor as compared to administration of either agent     alone, -   (2) provide for the administration of lesser amounts of the     administered chemotherapeutic agents, -   (3) provide for a chemotherapeutic treatment that is well tolerated     in the patient with fewer deleterious pharmacological complications     than observed with single agent chemotherapies and certain other     combined therapies, -   (4) provide for treating a broader spectrum of different cancer     types in mammals, especially humans, -   (5) provide for a higher response rate among treated patients, -   (6) provide for a longer survival time among treated patients     compared to standard chemotherapy treatments, -   (7) provide a longer time for tumor progression, and/or -   (8) yield efficacy and tolerability results at least as good as     those of the agents used alone, compared to known instances where     other cancer agent combinations produce antagonistic effects.

In one aspect of the invention, there is provided a method of treating type II diabetes, comprising administering a therapeutically effective amount of a compound according to the present invention, to a patient in need thereof.

In one aspect of the invention, there is provided a method of treating hypoxic tumors, comprising administering a therapeutically effective amount of a compound according to the present invention, to a patient in need thereof.

In one aspect of the invention, there is provided a method of treating a disease selected from inflammatory diseases, autoimmune diseases, neurodegenerative disorders, and viral infections, comprising administering a therapeutically effective amount of a compound according to the present invention, to a patient in need thereof.

In one aspect of the invention, there is provided a pharmaceutical composition comprising a compound according to the present invention, and a pharmaceutically acceptable diluent, carrier and/or excipient.

In one aspect of the invention, there is provided a pharmaceutical composition, comprising a therapeutically effective amount of a compound according to claim 1 and another anticancer agent selected from alkylating agents, antimetabolites, anticancer camptothecin derivatives, plant-derived anticancer agents, antibiotics, enzymes, platinum coordination complexes, tyrosine kinase inhibitors, hormones, hormone antagonists, monoclonal antibodies, interferons, and biological response modifiers.

As used herein, the term “C₁-C₆alkyl” means both linear and branched chain saturated hydrocarbon groups with 1 to 6 carbon atoms. Examples of C₁-C₆alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, t-butyl, n-pentyl, 4-methyl-butyl, n-hexyl, 2-ethyl-butyl groups. Among unbranched C₁-C₆alkyl groups, typical ones are methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl groups. Among branched alkyl groups, there may be mentioned iso-propyl, iso-butyl, sec-butyl, t-butyl, 4-methyl-butyl and 2-ethyl-butyl groups.

As used herein, the term “C₁-C₃alkyl” means both linear and branched chain saturated hydrocarbon groups with 1 to 3 carbon atoms. Examples of C₁-C₃alkyl groups include methyl, ethyl, n-propyl and isopropyl groups.

As used herein, the term “C₁-C₆alkoxy” means the group O—C₁-C₆alkyl, where “C₁-C₆alkyl” is used as described above. Examples of C₁-C₆alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, n-propoxy, n-butoxy, n-hexoxy, 3-methyl-butoxy groups.

As used herein, the term “C₁-C₃alkoxy” means the group O—C₁-C₃alkyl, where “C₁-C₃alkyl” is used as described above. Examples of C₁-C₃alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy and n-propoxy.

As used herein, the term “C₁-C₆haloalkyl” means both linear and branched chain saturated hydrocarbon groups, with 1 to 6 carbon atoms and with 1 to all hydrogens substituted by a halogen of different or same type. Examples of C₁-C₆haloalkyl groups include methyl substituted with 1 to 3 halogen atoms, ethyl substituted with 1 to 5 halogen atoms, n-propyl or iso-propyl substituted with 1 to 7 halogen atoms, n-butyl or iso-butyl substituted with 1 to 9 halogen atoms, and sec-butyl or t-butyl groups substituted with 1 to 9 halogen atoms.

As used herein, the term “C₁-C₃haloalkyl” means both linear and branched chain saturated hydrocarbon groups, with 1 to 3 carbon atoms and with 1 to all hydrogens substituted by a halogen of different or same type. Examples of C₁-C₃haloalkyl groups include methyl substituted with 1 to 3 halogen atoms, ethyl substituted with 1 to 5 halogen atoms, and n-propyl or iso-propyl substituted with 1 to 7 halogen atoms.

As used herein, the term “C₁-C₃haloalkoxy” means both linear and branched chain saturated alkoxy groups, with 1 to 3 carbon atoms and with 1 to all hydrogen atoms substituted by a halogen atom of different or same type. Examples of C₁-C₃haloalkoxy groups include methoxy substituted with 1 to 3 halogen atoms, ethoxy substituted with 1 to 5 halogen atoms, and n-propoxy or iso-propoxy substituted with 1 to 7 halogen atoms.

As used herein, the term “C₁-C₃fluoroalkyl” means both linear and branched chain saturated hydrocarbon groups, with 1 to 3 carbon atoms and with 1 to all hydrogen atoms substituted by a fluorine atom. Examples of C₁-C₃fluoroalkyl groups include methyl substituted with 1 to 3 fluorine atoms, ethyl substituted with 1 to 5 fluorine atoms, and n-propyl or iso-propyl substituted with 1 to 7 fluorine atoms.

As used herein, the term “C₁-C₃fluoroalkoxy” means both linear and branched chain saturated alkoxy groups, with 1 to 3 carbon atoms and with 1 to all hydrogen atoms substituted by a fluorine atom. Examples of C₁-C₃fluoroalkoxy groups include methoxy substituted with 1 to 3 fluorine atoms, ethoxy substituted with 1 to 5 fluorine atoms, and n-propoxy or iso-propoxy substituted with 1 to 7 fluorine atoms.

As used herein, the term “C₃-C₆cycloalkyl” means a cyclic saturated hydrocarbon group, with 3 to 6 carbon atoms. Examples of C₃-C₆cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, the term “C₁-C₃alkoxyC₁-C₃alkyl” means a both linear and branched chain saturated hydrocarbon group, with 1 to 3 carbon atoms, substituted with an alkoxy group with 1 to 3 carbon atoms. Examples of C₁-C₃alkoxyC₁-C₃alkyl groups are drawn below.

As used herein, the term “C₁-C₃cyanoalkyl” means both a linear and branched chain cyano (CN) derivative, with one to three carbon atoms including the carbon atom that is part of the cyano group. Examples of C₁-C₃cyanoalkyl groups are drawn below.

As used herein, the term N—C₁-C₃alkylamino means an amino substituent carrying one C₁-C₃alkyl group as defined supra. Examples of N—C₁-C₃alkylamino are drawn below.

As used herein, the term N,N-diC₁-C₃alkylamino means an amino substituent carrying two C₁-C₃alkyl groups as defined supra. Examples of N,N-diC₁-C₃alkylamino are drawn below.

As used herein, the term “halogen” means fluorine, chlorine, bromine or iodine. As used herein, the term “halo” means fluoro, chloro, bromo or iodo. It is to be understood that when a substituent is halo, it is always bound to a carbon.

As used herein, the term “heteroaryl” means a monocyclic aromatic group of carbon atoms wherein from one to three of the carbon atoms is/are replaced by one or more heteroatoms independently selected from nitrogen, oxygen or sulfur. In a bicyclic aryl, one of the rings may be partially saturated.

As used herein, the term “monocyclic heteroaryl” means a monocyclic aromatic group of carbon atoms wherein from one to three of the carbon atoms is/are replaced by one or more heteroatoms independently selected from nitrogen, oxygen or sulfur.

Examples of monocyclic heteroaryl groups include, but are not limited to, furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, pyridyl, triazolyl, triazinyl, pyridazyl, isothiazolyl, isoxazolyl, pyrazinyl, pyrazolyl, and pyrimidinyl.

As used herein, the term “heterocyclyl” means a cyclic group of carbon atoms wherein from one to three of the carbon atoms is/are replaced by one or more heteroatoms independently selected from nitrogen, oxygen and sulfur. Examples of heterocyclyl groups include, but are not limited to, tetrahydrofuryl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and dioxanyl.

Depending on the substituents present in compounds of the formula (I), the compounds may form salts which are within the scope of the present invention. Salts of compounds of formula (I), which are suitable for use in medicine are those wherein a counterion is pharmaceutically acceptable.

Suitable salts according to the invention include those formed with organic or inorganic acids or bases. In particular, suitable salts formed with acids according to the invention include those formed with mineral acids, strong organic carboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atoms which are unsubstituted or substituted, for example, by halogen, such as saturated or unsaturated dicarboxylic acids, such as hydroxycarboxylic acids, such as amino acids, or with organic sulfonic acids, such as (C₁-C₄)alkyl or aryl sulfonic acids which are unsubstituted or substituted, for example by halogen. Pharmaceutically acceptable acid addition salts include those formed from hydrochloric, hydrobromic, sulphuric, nitric, citric, tartaric, acetic, phosphoric, lactic, pyruvic, acetic, trifluoroacetic, succinic, perchloric, fumaric, maleic, glycolic, lactic, salicylic, oxaloacetic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, benzenesulfonic, isethionic, ascorbic, malic, phthalic, aspartic, and glutamic acids, lysine and arginine.

Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts, for example those of potassium and sodium, alkaline earth metal salts, for example those of calcium and magnesium, and salts with organic bases, for example dicyclohexylamine, N-methyl-D-glucamine, morpholine, thiomorpholine, piperidine, pyrrolidine, a mono, di- or tri lower alkylamine, for example ethyl, tertbutyl, diethyl, diisopropyl, triethyl, tributyl or dimethylpropylamine, or a mono-, di- or trihydroxy lower alkylamine, for example mono-, di- or triethanolamine. Corresponding internal salts may furthermore be formed.

The compounds of the invention may be used in the prophylaxis and/or treatment as such, or in a form of a pharmaceutical composition. While it is possible for the active ingredient to be administered alone, it is also possible for it to be present in a pharmaceutical composition. Accordingly, the invention provides a pharmaceutical composition comprising a compound of formula (I), and a pharmaceutically acceptable diluent, excipient and/or carrier. Pharmaceutical compositions of the invention may take the form of a pharmaceutical composition as described below.

Exemplary compositions for oral administration include suspensions which can contain, for example, microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or flavoring agents such as those known in the art; and immediate release tablets which can contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, calcium sulfate, sorbitol, glucose and/or lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants such as those known in the art. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Disintegrators include without limitation starch, methylcellulose, agar, bentonite, xanthan gum and the like. The compounds of formula (I) can also be delivered through the oral cavity by sublingual and/or buccal administration. Molded tablets, compressed tablets or freeze-dried tablets are exemplary forms which may be used. Exemplary compositions include those formulating the present compound(s) with fast dissolving diluents such as mannitol, lactose, sucrose and/or cyclodextrins. Also included in such compositions may be high molecular weight excipients such as celluloses (avicel) or polyethylene glycols (PEG). Such compositions can also include an excipient to aid mucosal adhesion such as hydroxy propyl cellulose (HPC), hydroxy propyl methyl cellulose (HPMC), sodium carboxy methyl cellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and agents to control release such as polyacrylic copolymer (e.g. Carbopol 934). Lubricants, glidants, flavors, coloring agents and stabilizers may also be added for ease of fabrication and use. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. For oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.

Compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets, pills or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid, for example as elixirs, tinctures, suspensions or syrups; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary or paste.

A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricating, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein. The present compounds can, for example, be administered in a form suitable for immediate release or extended release. Immediate release or extended release can be achieved by the use of suitable pharmaceutical compositions comprising the present compounds, or, particularly in the case of extended release, by the use of devices such as subcutaneous implants or osmotic pumps. The present compounds can also be administered liposomally.

Typical unit dosage compositions are those containing an effective dose, as hereinbefore recited, or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularly mentioned above, the compositions of this invention may include other agents conventional in the art having regard to the type of composition in question, for example those suitable for oral administration may include flavoring agents.

The compositions may be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Methods may include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. Compositions may be prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired composition.

The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, 1,2-dipalmitoylphosphatidylcholine, phosphatidyl ethanolamine (cephaline), phosphatidylserine, phosphatidylinositol, diphosphatidylglycerol (cardiolipin) or phosphatidylcholine (lecithin).

Compositions for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example saline or water-for-injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described. Exemplary compositions for parenteral administration include injectable solutions or suspensions which can contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as polyethylene glycol, ethanol, 1,3-butanediol, water, Ringer's solution, an isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid, or Cremaphor.

Exemplary compositions for nasal, aerosol or inhalation administration include solutions in saline, which can contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, and/or other solubilizing or dispersing agents such as those known in the art.

Compositions for rectal administration may be presented as a suppository with the usual carriers such as cocoa butter, synthetic glyceride esters or polyethylene glycol. Such carriers are typically solid at ordinary temperatures, but liquefy and/or dissolve in the rectal cavity to release the drug.

Compositions for topical administration in the mouth, for example buccally or sublingually, include lozenges comprising the active ingredient in a flavored basis such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a basis such as gelatin and glycerine or sucrose and acacia. Exemplary compositions for topical administration include a topical carrier such as Plastibase (mineral oil gelled with polyethylene).

Compounds of formula (I) may be administered as the sole pharmaceutical agent or in combination with one or more additional therapeutic agents where the combination causes no unacceptable adverse effects. This pharmaceutical composition includes administration of a single pharmaceutical dosage composition which contains a compound of formula (I) and one or more additional therapeutic agents, as well as administration of the compound of formula (I) and each additional therapeutic agent in its own separate pharmaceutical dosage composition. For example, a compound of formula (I) and a therapeutic agent may be administered to the patient together in a single oral dosage composition such as a capsule or tablet, or each agent may be administered in compositions with separate dosage.

Where separate dosage compositions are used, the compound of formula (I) and one or more additional therapeutic agents may be administered at essentially the same time (e.g., concurrently) or at separately staggered times (e.g., sequentially).

The amount of active ingredient which is required to achieve a therapeutic effect will, of course, vary with the particular compound, the route of administration, the subject under treatment, including the type, species, age, weight, sex, and medical condition of the subject and the renal and hepatic function of the subject, and the particular disorder or disease being treated, as well as its severity. An ordinarily skilled physician, veterinarian or clinician can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.

Oral dosages of the present invention, when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day, preferably 0.01 mg per kg of body weight per day (mg/kg/day) to 10 mg/kg/day, and most preferably 0.1 to 5.0 mg/kg/day, for adult humans. For oral administration, the compositions may be provided in the form of tablets or other forms of presentation provided in discrete units containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, and milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from about 1 mg to about 100 mg of active ingredient. Intravenously, the most preferred doses will range from about 0.1 to about 10 mg/kg/minute during a constant rate infusion. Compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in the art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.

Preparation of Compounds

The compounds in the present invention can be prepared as a free base or a pharmaceutically acceptable salt thereof by the process described below. Throughout the following description of such processes it is understood that, where appropriate, suitable protecting groups will be added to, and subsequently removed from the various reactants and intermediates in a manner that will be readily understood by one skilled in the art of organic synthesis. Conventional procedures for using such protecting groups as well as examples of suitable protecting groups are for example described in Protective Groups in Organic Synthesis by T. W. Greene, P. G. M Wutz, 4^(th) Edition, Wiley-Interscience, New York, 2006. It is understood that microwaves can alternatively be used for the heating of reaction mixtures.

Another aspect of the present invention provides a process for preparing a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein R¹, R² and A are, unless specified otherwise, as defined herein. Said process comprises of:

(i) Formation of a Corresponding Compound of Formula (I)

A compound of formula (I) may be obtained (Scheme 1) by starting from, for example, a compound of formula (II), wherein R^(X) may be F, OCH₃, OC(CH₃)₃, or OSiR′R″R′″ (wherein R′, R″ and R′″ are independently aryl (such as phenyl) or alkyl (such as methyl or tert-butyl)). If R^(X) is F the conversion into (I) may be carried out by for instance acidic hydrolysis using aqueous HCl. If R^(X) is OCH₃ the conversion into (I) may be carried out by reaction with for instance trimethylsilyl iodide in a suitable solvent such as chloroform or by reaction with HBr in a suitable solvent such as acetic acid or by reaction with BBr₃ in a suitable solvent such as dichloromethane. If R^(X) is OC(CH₃)₃ the conversion into (I) may be carried out by reaction with for instance trifluoroacetic acid in a suitable solvent such as dichloromethane. If R^(X) is OSiR′R″R′″ the conversion into (I) may be carried out by for instance HCl in a suitable solvent such as methanol or by using tetrabutyl ammonium fluoride in tetrahydrofuran. If enantiomerically pure or enriched compound (II) is used in this reaction, an enantiomerically pure or enantiomerically enriched compound (I) is obtained.

Compounds of formula (II) are commercially available compounds, or are known in the literature, or they are prepared by standard processes known in the art. A compound of formula (I) or (II) may be separated into its enantiomers by standard processes known in the art by for example chromatography on a chiral stationary phase.

General Methods

All solvents used were of analytical grade and commercially available anhydrous solvents were routinely used for reactions. Starting materials were available from commercial sources, or prepared according to literature procedures. Room temperature refers to +20-25° C. Solvent mixture compositions are given as volume percentages or volume ratios.

Microwave heating was performed in a Biotage Initiator microwave cavity producing continuous irradiation at 2.45 GHz. It is understood that microwaves may be used for the heating of reaction mixtures.

Straight phase chromatography was manually performed on Merck Silica gel (0.040-0.063 mm), or automatically using an ISCO Combiflash® Companion™ system using SiliaSep™ normal-phase flash columns using the solvent system indicated.

NMR spectra were recorded on a 400 MHz (or higher field) NMR spectrometer fitted with a probe of suitable configuration. Spectra were recorded at ambient temperature unless otherwise stated. Chemical shifts are given in ppm down- and upfield from TMS (0.00 ppm). The following reference signals were used: the residual solvent signal of DMSO-d₆ δ 2.5, CDCl₃ δ 7.26 or Methanol-d₄ δ 3.31. Resonance multiplicities are denoted s, d, t, q, m and br for singlet, doublet, triplet, quartet, multiplet and broad, respectively.

High pressure liquid chromatography (HPLC) was performed on a reverse phase column. A linear gradient was applied using for example mobile phase A (aqueous 0.1% NH₃ or aqueous 0.1% acetic acid or aqueous 0.1% formic acid) and B (acetonitrile or methanol). Mass spectrometer (MS) analyses were performed in positive ion mode using electrospray ionization (ES+).

Preparative chromatography was run on a Gilson-PREP GX271 or GX281 with Trilution Ic as software on a reverse phase column. A linear gradient was applied using for example mobile phase A (aqueous 0.1% NH₃ or aqueous 0.1% acetic acid or aqueous 0.1% formic acid) and B (acetonitrile or methanol).

Preparative chiral chromatography for separation of enantiomers was run on a Thar SFC using supercritical fluid chromatography on a chiral stationary phase. A linear gradient was applied using mobile phase A (carbon dioxide) and B (acetonitrile or methanol or ethanol or 2-propanol or any mixtures thereof). Additives (such as diethyl amine or isopropyl amine or ammonia or formic acid or TFA) may be used.

Compounds have been named using BIOVIA Draw 16.1.

Abbreviations

-   Amphos (4-(N,N-Dimethylamino)phenyl)di-tert-butyl phosphine -   anh. anhydrous -   aq. aqueous -   BuLi butyl lithium -   DCM dichloromethane -   DMAc N,N-dimethyl acetamide -   DME 1,2-Dimethoxyethane -   DMF N,N-dimethyl formamide -   DMSO dimethyl sulfoxide -   EtOAc ethyl acetate -   EtOH ethanol -   h hour(s) -   HPLC high pressure (or performance) liquid chromatography -   KOtBu potassium tert-butoxide -   LCMS liquid chromatography mass spectrometry -   MeCN acetonitrile -   2-MeTHF 2-methyl tetrahydrofuran -   MeOH methanol -   min. minute(s) -   NMR nuclear magnetic resonance -   PEPPSI-iPr     [1,3-Bis(2,6-Diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II)     dichloride -   Pd(OAc)₂ palladium(II) acetate -   PdCl₂(dppf)     [1,1′-Bis(diphenylphosphino)ferrocene]-dichloropalladium(II) -   quant. quantitative -   rt room temperature -   sat. saturated -   TFA trifluoroacetic acid -   THF tetrahydrofuran

Example 1 (3R)-4-(2,6-dichloro-4-pyridyl)-3-methyl-morpholine

2,6-Dichloro-4-iodo-pyridine (2 g, 7.3 mmol), (3R)-3-methylmorpholine hydrochloride (1.1 g, 7.99 mmol), PPh₃ (120 mg, 0.46 mmol), Pd(OAc)₂ (50 mg, 0.22 mmol) and freshly ground K₃PO₄ (5.5 g, 25.9 mmol) were taken up in DMF (15 ml). The resulting mixture was degassed with nitrogen for 5 min and stirred at 100° C. overnight. When cooled to rt the mixture was poured into water (50 ml) and EtOAc (15 ml). The organic layer was separated and the aqueous layer was extracted with EtOAc (3×10 ml). The combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 10-60% EtOAc in heptane to give the product as a solid (520 mg, 29%). MS ES+ m/z 247 [M+H]⁺.

Example 2 (3R)-4-(2-tert-butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine

(3R)-4-(2,6-dichloro-4-pyridyl)-3-methyl-morpholine (520 mg, 2.1 mmol), KOtBu (500 mg, 4.46 mmol) and 4 Å molecular sieves (˜10 beads, 4-8 mesh) were taken up in toluene (8 ml) and stirred at 90° C. for 2 h. When cooled to rt the mixture was diluted with EtOAc (10 ml), brine (5 ml) and water (5 ml). The organic layer was separated and the aqueous layer extracted with EtOAc (10 ml). The combined organics were washed with brine, concentrated and purified on a silica gel column eluted with 0-40% EtOAc in heptane to give the product as an oil. MS ES+ m/z 285 [M+H]⁺.

Example 3 tert-Butyl 4-[6-tert-butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-3-(trifluoromethyl)piperazine−1-carboxylate

A mixture of tert-butyl 3-(trifluoromethyl)piperazine-1-carboxylate (500 mg, 1.97 mmol), (3R)-4-(2-tert-butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine (670 mg, 2.36 mmol), KOtBu (440 mg, 3.94 mmol) and PEPPSI-Ipr (133 mg, 0.2 mmol) in 1,4-dioxane (15 ml) was heated in a microwave reactor at 130° C. for 3 h. The reaction mixture was poured into water (10 ml) and extracted with EtOAc (3×20 ml). The combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 20-30% EtOAc in heptane to give the product as an oil (420 mg, 42%). MS ES+ m/z 503 [M+H]⁺.

Example 4 (3R)-4-[2-tert-butoxy-6-[2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

LiAlH₄ (63 mg, 1.67 mmol) was added at 0° C. to a solution of tert-butyl 4-[6-tert-butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-3-(trifluoromethyl)piperazine-1-carboxylate (420 mg, 0.84 mmol) in THF (10 ml) and the resulting mixture was stirred at rt for 5 h. The reaction mixture was quenched with ice/water and extracted with EtOAc (3×10 ml). The combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 4-5% MeOH in DCM to give the product as a solid (200 mg, 59%). MS ES+ m/z 403 [M+H]⁺.

Example 5 (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

Methanesulfonyl chloride (85 mg, 0.75 mmol) was added drop wise to a solution of (3R)-4-[2-tert-butoxy-6-[2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine (200 mg, 0.5 mmol) and Et₃N (100 mg, 0.99 mmol) in DCM (15 ml) at 0° C. and the resulting mixture was stirred at rt for 3 h. The reaction mixture was quenched with ice/water and extracted with EtOAc (3×10 mL). The combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 0-100% EtOAc in heptane to give the product as a solid (150 mg, 63%). MS ES+ m/z 481 [M+H]⁺.

Example 6 (R) and (S) 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

TFA (0.4 ml, 5.23 mmol) was added to a solution of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine (150 mg, 0.31 mmol) in DCM (15 ml) at 0° C. and the resulting mixture was stirred at rt for 6 h. The pH was adjusted above 7 using sat. aq. NaHCO₃ and the mixture extracted with DCM (3×10 ml). The combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 0-10% MeOH in DCM to give the product as a solid (20 mg, 23%).

Chiral separation by SFC gave the two isomers.

Example 6-1, First Isomer to Elute, with Unknown Absolute Configuration

¹H NMR (400 MHz, DMSO-d₆):δ 10.45 (br.s., 1H), 5.75 (br.s., 1H), 5.56 (br. S., 1H), 5.43 (br.s., 1H), 4.09 (br.s., 1H), 3.90-3.69 (m, 3H), 3.66-3.35 (m, 2H), 3.51-3.39 (m 3H), 3.21-3.10 (m, 1H), 3.10-3.08 (m, 1H), 3.02-2.69 (m, 1H), 2.95-2.91 (s, 3H), 2.84-2.79 (m, 1H), 1.06-1.05 (m, 3H). MS ES+ m/z 425 [M+H]⁺.

Example 6-2, Second Isomer to Elute, with Unknown Absolute Configuration

¹H NMR (400 MHz, DMSO-d₆):δ 10.45 (br.s., 1H), 5.76 (br.s., 1H), 5.56 (br.s., 1H), 5.43 (br.s., 1H), 4.09 (br.s., 1H), 3.90-3.69 (m, 3H), 3.66-3.35 (m, 2H), 3.51-3.39 (m 3H), 3.21-3.10 (m, 1H), 3.10-3.08 (m, 1H), 3.02-2.69 (m, 1H), 2.95-2.91 (s, 3H), 2.84-2.79 (m, 1H), 1.06-1.05 (m, 3H). MS ES+ m/z 425 [M+H]⁺.

Example 7 (3R)-4-[2-tert-butoxy-6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using (4-fluorophenyl)methanesulfonyl chloride instead of methanesulfonyl chloride, to give the product as a solid (40 mg, 37%). MS ES+ m/z 575 [M+H]⁺.

Example 8 6-[4-[(4-Fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (12 mg, 33%). ¹H NMR (400 MHz, DMSO-d₆):δ 7.4 (q, 2H), 7.2 (t, 2H), 5.7 (br.s., 1H), 5.43 (br.s., 1H), 4.5 (br.s., 2H), 4.09 (t, 1H), 3.90 (t, 2H), 3.8 (d, 1H), 3.6 (q, 3H), 3.4 (m, 3H), 3.08 (m, 3H), 2.8 (t, 1H), 1.06-1.05 (t, 3H). MS ES+ m/z 519 [M+H]⁺.

Example 9 (3R)-4-[2-tert-butoxy-6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using 5-fluoropyridine-3-sulfonyl chloride instead of methanesulfonyl chloride, to give the product as a solid (40 mg, 32%). MS ES+ m/z 562 [M+H]⁺.

Example 10 6-[4-[(5-Fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (26 mg, 72%). ¹H NMR (400 MHz, DMSO-d₆):δ 9.8 (br.s., 1H), 8.9 (d, 1H), 8.8 (br.s., 1H), 8.2 (d, 1H), 5.7 (br.s., 1H), 5.6 (s, 1H), 5.5 (s, 1H), 5.4 (s, 1H), 4.0 (t, 2H), 3.8 (d, 2H), 3.7-3.6 (t, 2H), 3.5 (d, 1H), 3.4 (m, 1H), 3.3 (br.s., 1H), 3.2 (d, 1H), 2.9 (m, 1H), 2.8 (d, 1H), 2.5 (m, 1H), 1.2 (t, 1H), 1.06-1.05 (q, 3H). MS ES+ m/z 506 [M+H]⁺.

Example 11 (3R)-4-[2-tert-butoxy-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using tetrahydrofuran-3-sulfonyl chloride instead of methanesulfonyl chloride, to give the product as a solid (35 mg, 35%). MS ES+ m/z 537 [M+H]⁺.

Example 12 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (26 mg, 83%). ¹H NMR (400 MHz, DMSO-d₆): δ 9.8 (br.s., 1H), 5.7 (d, 1H), 5.5 (d, 1H), 4.0 (m, 2H), 3.9-3.7 (m, 6H), 3.6 (m, 4H), 3.5 (m, 3H), 3.0 (m, 2H), 2.4-2.1 (m, 1H), 2.0 (m, 1H), 1.9-1.1 (q, 1H), 1.0 (q, 3H). MS ES+ m/z 481 [M+H]⁺.

Example 13 (3R)-4-[2-tert-butoxy-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using pyrrolidine-1-sulfonyl chloride instead of methanesulfonyl chloride, to give the product as a solid (76 mg, 71%). MS ES+ m/z 536 [M+H]⁺.

Example 14 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (48 mg, 72%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.05-1.12 (m, 3H), 1.82-1.89 (m, 4H), 2.85 (br t, 1H), 3.00 (td, 1H), 3.10 (br d, 1H), 3.16-3.31 (m, 5H), 3.33-3.42 (m, 1H), 3.44-3.56 (m, 2H), 3.59-3.65 (m, 1H), 3.66-3.71 (m, 1H), 3.83 (dd, 1H), 3.90 (br dd, 2H), 4.01 (br s, 1H), 5.43 (br s, 1H), 5.49 (br s, 1H), 5.70 (br s, 1H), 9.81 (br s, 1H). MS ES+ m/z 480 [M+H]⁺.

Example 15 4-[6-tert-Butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-N,N-dimethyl-3-(trifluoromethyl)piperazine-1-sulfonamide

The title compound was prepared as described in Example 5, using N,N-dimethylsulfamoyl chloride instead of methanesulfonyl chloride, to give the product as a solid (60 mg, 59%). MS ES+ m/z 510 [M+H]⁺.

Example 16 N,N-dimethyl-4-[4-[(3R)-3-methylmorpholin-4-yl]-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide

The title compound was prepared as described in Example 6 using 4-[6-tert-Butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-N,N-dimethyl-3-(trifluoromethyl)piperazine-1-sulfonamide instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (18 mg, 37%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.05-1.12 (m, 3H), 2.80 (d, 6H), 2.86-2.95 (m, 1H), 3.00 (td, 1H), 3.12-3.24 (m, 2H), 3.34-3.41 (m, 1H), 3.44-3.56 (m, 2H), 3.59-3.64 (m, 1H), 3.66-3.74 (m, 1H), 3.81-3.95 (m, 3H), 4.04 (br s, 1H), 5.44 (br s, 1H), 5.49 (br s, 1H), 5.70 (br s, 1H), 9.50-10.12 (m, 1H). MS ES+m/z 454 [M+H]⁺.

Example 17 (3R)-4-[2-tert-butoxy-6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using 2-methoxyethanesulfonyl chloride instead of methanesulfonyl chloride, to give the product as a solid (74 mg, 76%). MS ES+ m/z 525 [M+H]⁺.

Example 18 6-[4-(2-Methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (48 mg, 75%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.05-1.16 (m, 3H), 2.90 (br t, 1H), 3.00 (td, 1H), 3.11-3.23 (m, 2H), 3.25-3.30 (m, 3H), 3.33-3.51 (m, 4H), 3.55-3.71 (m, 5H), 3.83-3.96 (m, 3H), 4.05 (br s, 1H), 5.44 (br s, 1H), 5.50 (br s, 1H), 5.72 (br s, 1H), 9.81 (br s, 1H). MS ES+ m/z 469 [M+H]⁺.

Example 19 (3R)-4-[2-tert-butoxy-6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using 4-fluorobenzenesulfonyl chloride instead of Methanesulfonyl chloride, to give the product as a solid (40 mg, 41%). MS ES+ m/z 561 [M+H]⁺.

Example 20 6-[4-(4-Fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (22 mg, 61%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.04 (dd, 3H), 2.28-2.36 (m, 1H), 2.53-2.60 (m, 1H), 2.97 (td, 1H), 3.23 (br t, 1H), 3.40-3.54 (m, 3H), 3.55-3.72 (m, 3H), 3.84-4.01 (m, 2H), 4.56 (t, 1H), 5.40 (br s, 1H), 5.49 (br s, 1H), 5.67 (br s, 1H), 7.47-7.53 (m, 2H), 7.86 (dd, 2H), 9.78 (br s, 1H). MS ES+ m/z 505 [M+H]⁺.

Example 21 (3R)-4-[2-tert-butoxy-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using 2-methylpyrazole-3-sulfonyl chloride instead of methanesulfonyl chloride, to give the product as a solid (90 mg, 82%). MS ES+ m/z 547 [M+H]⁺.

Example 22 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine, to give the product as a solid (35 mg, 54%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.06 (dd, 3H), 2.64-2.74 (m, 1H), 2.92-3.03 (m, 2H), 3.20-3.30 (m, 1H), 3.33-3.52 (m, 2H), 3.57-3.64 (m, 1H), 3.65-3.74 (m, 2H), 3.81-3.91 (m, 2H), 3.93-4.02 (m, 1H), 4.03-4.06 (m, 3H), 4.06-4.14 (m, 1H), 5.44 (s, 1H), 5.55 (br s, 1H), 5.71 (s, 1H), 6.92 (dd, 1H), 7.65 (dd, 1H), 9.82 (br s, 1H). MS ES+ m/z 491 [M+H]⁺.

Example 23 (3R)-4-[2-tert-butoxy-6-[4-cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using cyclopropanesulfonyl chloride, to give the product as a solid (80 mg, 79%). MS ES+ m/z 507 [M+H]⁺.

Example 24 6-[4-Cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, to give the product as a solid (42 mg, 59%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.93-1.09 (m, 7H), 2.53-2.65 (m, 1H), 2.90-3.05 (m, 2H), 3.16-3.24 (m, 2H), 3.33-3.52 (m, 2H), 3.56-3.64 (m, 2H), 3.65-3.72 (m, 1H), 3.85-3.96 (m, 3H), 4.10 (br s, 1H), 5.44 (br s, 1H), 5.56 (br d, 1H), 5.76 (s, 1H), 9.77 (br s, 1H). MS ES+ m/z 451 [M+H]⁺.

Example 25 (3R)-4-[2-tert-butoxy-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepare as described in Example 5, using piperidine-1-sulfonyl chloride, to give the product as a solid (100 mg, 92%). MS ES+ m/z 550 [M+H]⁺.

Example 26 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, to give the product as a solid (42 mg, 47%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.07 (dd, 3H), 1.52 (br s, 6H), 2.85-3.03 (m, 1H), 3.08-3.12 (m, 1H), 3.17 (br d, 5H), 3.34-3.39 (m, 1H), 3.45 (br s, 1H), 3.47-3.52 (m, 2H), 3.61 (dt, 1H), 3.69 (dd, 1H), 3.80 (d, 1H), 3.83 (d, 1H), 3.88-3.93 (m, 2H), 5.43 (s, 1H), 5.48 (br s, 1H), 5.70 (s, 1H), 9.32-9.88 (m, 1H). MS ES+ m/z 494 [M+H]⁺.

Example 27 (3R)-4-[2-tert-butoxy-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using morpholine-4-sulfonyl chloride, to give the product as a solid (75 mg, 77%). MS ES+ m/z 483 [M+H]⁺.

Example 28 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, to give the product as a solid (16 mg, 64%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.07 (dd, 3H), 2.88-3.08 (m, 2H), 3.12-3.23 (m, 6H), 3.36-3.41 (m, 1H), 3.43-3.50 (m, 1H), 3.52-3.70 (m, 7H), 3.82-3.96 (m, 3H), 4.02 (br s, 1H), 5.44 (br s, 1H), 5.49 (br s, 1H), 5.71 (br s, 1H), 9.84 (br s, 1H). MS ES+m/z 496 [M+H]⁺.

Example 29 (3R)-4-[2-tert-butoxy-6-[4-(1,2-dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5, using 1,2-dimethylimidazole-4-sulfonyl chloride, to give the product as a solid (120 mg, 86%). MS ES+ m/z 561 [M+H]⁺.

Example 30 6-[4-(1,2-Dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, to give the product as a solid (24 mg, 23%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.04-1.09 (m, 3H), 2.25-2.32 (m, 3H), 2.41-2.49 (m, 1H), 2.65-2.82 (m, 1H), 2.91-3.06 (m, 1H), 3.09-3.29 (m, 1H), 3.29-3.49 (m, 3H), 3.59-3.62 (m, 1H), 3.64-3.71 (m, 1H), 3.82-3.94 (m, 4H), 4.08 (br d, 1H), 4.56-4.58 (m, 1H), 5.41 (s, 1H), 5.47 (br d, 1H), 5.64-5.73 (m, 1H), 7.62-7.84 (m, 1H), 7.80 (s, 1H), 10.45 (m, 1H). MS ES+ m/z 505 [M+H]⁺.

Example 31 6-[4-(1-methylcyclopropyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

To an ice cooled solution of (3R)-4-[2-tert-butoxy-6-[2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine (70 mg, 0.17 mmol in DCM (5 ml) was added TEA (0.12 ml, 0.87 mmol) and 1-methylcyclopropanesulfonyl chloride (40 mg, 0.26 mmol). The cooling bath was removed, and the resulting mixture was stirred at rt overnight. More 1-methylcyclopropanesulfonyl chloride (40 mg, 0.26 mmol) was added and the reaction was stirred at rt for 20 h. More 1-methylcyclopropanesulfonyl chloride (40 mg, 0.26 mmol) was added and the reaction was kept at rt for 2 months. The resulting residue was purified by preparative HPLC to give the product as a solid (21 mg, 26%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.80-0.90 (m, 2H), 1.06-1.07 (m, 3H) 1.12-1.28 (m, 2H), 1.41 (s, 3H), 2.94-3.10 (m, 2H), 3.12-3.24 (m, 1H), 3.28-3.31 (m, 1H), 3.35-3.41 (m, 1H), 3.42-3.51 (m, 1H), 3.58-3.73 (m, 3H), 3.87-4.00 (m, 3H), 4.03 (br d, 1H), 5.44 (br s, 1H), 5.48 (br s, 1H), 5.71 (s, 1H), 9.86 (br s, 1H). MS ES+ m/z 465 [M+H]⁺.

Example 32 (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-4-pyridyl]-3-methyl-morpholine

(3R)-4-(2-tert-butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine (85 mg, 0.3 mmol), K₂CO₃ (83 mg, 0.6 mmol), [4-methylsulfonyl-2-(trifluoromethyl)phenyl]boronic acid (80 mg, 0.3 mmol) and PdCl₂(Amphos) (11 mg, 0.02 mmol) were taken up in 1,4-dioxane (2 ml) and water (0.6 ml) and the resulting mixture was stirred at 95° C. for 1 h. The reaction allowed to cool and [4-methylsulfonyl-2-(trifluoromethyl)phenyl]boronic acid (161 mg, 0.6 mmol) and PdCl₂(Amphos) (11 mg, 0.02 mmol) were added and the reaction was heated again and stirred at 95° C. for 3 h. When cooled to rt the mixture was extracted with EtOAc and the combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 0-50% EtOAc in heptane to give the product as a solid (22 mg, 16%). MS ES+ m/z 473 [M+H]⁺.

Example 33 4-[(3R)-3-methylmorpholin-4-yl]-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, to give the product as a solid (8 mg, 47%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.10 (d, 3H), 3.03 (td, 1H), 3.30-3.39 (m, 1H), 3.33-3.41 (m, 3H), 3.43-3.54 (m, 1H), 3.58-3.70 (m, 2H), 3.86-3.95 (m, 2H), 5.48 (br s, 1H), 6.08 (br s, 1H), 7.87 (d, 1H), 8.28-8.33 (m, 2H), 11.07 (br s, 1H). MS ES+ m/z 417 [M+H]⁺.

Example 34 4-[6-tert-Butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-N,N-dimethyl-3-(trifluoromethyl)benzenesulfonamide

A mixture of 4-bromo-N,N-dimethyl-3-(trifluoromethyl)benzenesulfonamide (199 mg, 0.6 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (183 mg, 0.72 mmol), KOAc (118 mg, 1.2 mmol) and PdCl₂(dppf) (44 mg, 0.06 mmol) in DMSO (2.5 ml) was stirred at 90° C. overnight. When cooled to rt, (3R)-4-(2-tert-butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine (171 mg, 0.6 mmol), K₂CO₃ (166 mg, 1.20 mmol), PdCl₂(dppf) (22 mg, 0.03 mmol), 1,4-dioxane (1 ml) and water (1 ml) were added and the reaction was stirred at 85° C. for 2 h. When cooled to rt, [4-(dimethylsulfamoyl)-2-(trifluoromethyl)phenyl]boronic acid (178 mg, 0.6 mmol), K₂CO₃ (83 mg, 0.6 mmol) and PdCl₂(dppf) (22 mg, 0.03 mmol) were added and the reaction was stirred at 85° C. for 5 h. The reaction was added to water and extracted with EtOAc. The combined organics were washed with brine, dried over Na₂SO₄, filtered, concentrated and purified on a silica gel column eluted with 0-80% EtOAc in heptane to give the product as a solid (200 mg, 67%). MS ES+ m/z 502 [M+H]⁺.

Example 35 N,N-dimethyl-4-[4-[(3R)-3-methylmorpholin-4-yl]-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)benzenesulfonamide

The title compound was prepared as described in Example 6. When the reaction was complete the mixture was concentrated, chased with toluene and purified by preparative HPLC to give the product as a solid (85 mg, 50%). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.10 (d, 3H), 2.66-2.78 (m, 6H), 3.03 (td, 1H), 3.38 (br s, 1H), 3.48 (td, 1H), 3.57-3.70 (m, 2H), 3.87-3.96 (m, 2H), 5.48 (br s, 1H), 6.12 (br s, 1H), 7.86 (d, 1H), 8.03 (d, 1H), 8.12 (dd, 1H), 11.02 (br s, 1H). MS ES+ m/z 446 [M+H]⁺.

Example 36 Vps34 Biochemical Assay

Dilution series of compounds of the invention were prepared in DMSO at 100 times the final assay concentration (n₁=n₀/3 in 10 points). The compounds were further diluted to 4 times the assay concentration in assay buffer (Life technologies buffer Q, PV5125, diluted 5 times supplemented with 2 mM DTT and 2 mM MnCl₂). 2.5 μl of the diluted compounds were added to a 384 well assay plate followed by 2.5 μl of 16.5 nM Vps34 enzyme (Life technologies, PV5126). Enzyme and compounds were pre-incubated at rt for 15 min. Then 5 μl of substrate mix containing 20 μM ATP (Life technologies, PV3227) and 200 μM PI:PS substrate (Life technologies, PV5122) in assay buffer was added to the wells containing compound and enzyme. Mixing was performed by pipetting several times. The reaction was incubated at room temperature for 1 h. Then 5 μl stop-detection mix, prepared as described in the Adapta kinase assay kit instructions (Life technologies, PV5099) containing Adapta Eu-anti-ADP antibody (2.3 nM), Alexa Fluor 647 ADP tracer (9 nM) and EDTA (30 mM) in TR-FRET buffer, was added to quench the reaction. Mixing was performed by pipetting several times. The assay plate was then incubated at room temperature for 30 min and read with Artemis micro plate reader. Percent inhibition of the compounds as compared to DMSO treated control samples was calculated. By the use of Dotmatics software compound concentration versus percent inhibition was fitted to generate IC₅₀ values.

The example compounds effectively inhibited Vps34 and the results of the assay are shown in Table 1 (Median IC₅₀ nM Adapta).

TABLE 1 Median IC₅₀ values for the Vps34 assay Example Median IC₅₀ Compound nM Adapta 6-1 <5 6-2 15 12 <5 10 <5  8 <5 16 <5 14 <5 18 <5 20 <5 22 <5 24 <5 26 <5 28 <5 30 <5 31 <5 33 <5 35 <5

Example 37

High Content Screening Autophagy assay Human osteosarcoma cells (HOS) stably expressing a Green Fluorescent Protein (GFP) tagged LC3 (GFP-LC3) were used to determine the inhibitory effect on autophagy of proprietary compounds. For that purpose, autophagy was activated by using the mTOR inhibitor KU-0063794 at 500 nM in the presence of Bafilomycin A1 (Sigma-Aldrich) at 5 nM. Shortly, cells were plated overnight in clear bottom 96-well plates in DMEM-High Modified media (Hi-Clone Cat #SH30285.01). At the start of the experiment, the media was removed and replaced with fresh media containing the mTOR inhibitor, Bafilomycin A1 and the vehicle or a test compound as indicated. After 6 hours the media was removed, cells were washed twice with ice-cold phosphate buffered saline (PBS) and fixed with 4% paraformaldehyde for 20 minutes at room temperature. Then the cells were washed twice with ice-cold PBS before adding Hoechst 33342 at 1 μg/ml in PBS for nuclear staining. After incubation overnight at 4° C., cells were washed once with PBS to remove the excess of dye and 100 μl of PBS was added to each well. Images were acquired at 20× magnification, 6 images per well, using the ImageXpress automated microscope (Molecular Devices Inc.) and analyzed with MetaXpress software to identify LC3-GFP foci. Foci area per cell values were used to generate dose response curves and IC50 values were calculated using the non-linear fitting analysis in GraphPad Prism software.

The tested example compounds effectively inhibited autophagy in HOS cells. The results of the assay are shown in Table 2 (Median IC₅₀ nM HOS-LC3).

TABLE 2 Median IC₅₀ values for the Vps34 assay and autophagy in HOS cells assay. Median IC₅₀ nM Example Cellular Compound assay 6-1 33 12 55 10 106  8 12 16 17 18 83 20 223 22 95 24 28 26 139 28 248 30 84 33 491 35 458 

1-44. (canceled)
 45. A method of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound represented by formula (I),

or a pharmaceutically acceptable salt thereof, wherein R¹ is C₁-C₃alkyl or cyclopropyl; R² is selected from hydrogen, C₁-C₃haloalkyl and C₁-C₃alkyl; A is

wherein R³ is selected from R⁶, C₁-C₆alkyl, amino, N—C₁-C₃alkylamino, N,N-diC₁-C₃alkylamino and C₁-C₃alkoxyC₁-C₃alkyl, said C₁-C₆alkyl and said C₁-C₃alkoxyC₁-C₃alkyl being optionally substituted with one R⁶ and/or one or more halo; R⁴ is selected from C₁-C₆alkyl, C₁-C₆alkoxy, C₁-C₆haloalkyl, C₃-C₆cycloalkyl and phenyl, said phenyl being optionally and independently substituted with one or more of fluoro, chloro, methyl, methoxy, dimethylamino, trifluoromethoxy, trifluoromethyl, and cyclopropyl; R⁵ is selected from halogen, C₁-C₆alkyl, C₁-C₆alkoxy, C₁-C₆haloalkyl and C₃-C₆cycloalkyl; R⁶ is selected from phenyl, monocyclic heteroaryl, C₃-C₆cycloalkyl, and heterocyclyl, each optionally substituted with one or more R⁷; and R⁷ is selected from halogen, amino, N—C₁-C₃alkylamino, N,N-diC₁-C₃alkylamino, C₁-C₃alkoxyC₁-C₃alkyl, C₁-C₃alkoxy, C₁-C₃haloalkoxy, C₃-C₆cycloalkyl, C₁-C₃haloalkyl and C₁-C₃alkyl.
 46. The method of claim 45, wherein the cancer is selected from the group consisting of breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, renal cancer, colon cancer, glioma, prostate cancer, ovarian cancer, melanoma, and lung cancer.
 47. The method of claim 45, further comprising administering to the patient radiation therapy.
 48. The method of claim 45, wherein R² is selected from hydrogen and C₁-C₃alkyl, or a pharmaceutically acceptable salt thereof.
 49. The method of claim 45, wherein R⁷ is selected from fluoro, cyclopropyl and methyl, or a pharmaceutically acceptable salt thereof.
 50. The method of claim 45, wherein R⁴ is selected from methyl, trifluoromethyl, cyclopropyl and phenyl, said phenyl being optionally meta-substituted with one of fluoro, chloro, methyl, methoxy, dimethylamino, trifluoromethoxy, trifluoromethyl and cyclopropyl; and R⁵ is selected from chloro, cyclopropyl, methyl and trifluoromethyl, or a pharmaceutically acceptable salt thereof.
 51. The method of claim 45, wherein R⁴ is selected from methyl, trifluoromethyl and cyclopropyl; and R⁵ is selected from chloro, cyclopropyl, methyl and trifluoromethyl, or a pharmaceutically acceptable salt thereof.
 52. The method of claim 45, wherein R³ is selected from R⁶, C₁-C₃alkyl, N,N-diC₁-C₃alkylamino and methoxyC₁-C₃alkyl, said C₁-C₃alkyl being optionally substituted with one R⁶, or a pharmaceutically acceptable salt thereof.
 53. The method of claim 45, wherein R⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, isoxazolyl, pyrazolyl, oxazolyl, cyclopropyl, cyclopentyl, pyrrolidinyl and tetrahydrofuryl, each optionally substituted with one or more R⁷, or a pharmaceutically acceptable salt thereof.
 54. The method of claim 45, wherein R⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, pyrazolyl, cyclopropyl, pyrrolidinyl, piperidinyl, and tetrahydrofuryl, each optionally substituted with one or more R⁷, or a pharmaceutically acceptable salt thereof.
 55. The method of claim 45, wherein R⁶ is selected from

or a pharmaceutically acceptable salt thereof.
 56. The method of claim 45, wherein R³ is selected from

or a pharmaceutically acceptable salt thereof.
 57. The method of claim 45, wherein R¹ is methyl or cyclopropyl; R² is hydrogen; A is

and R³ is selected from

or a pharmaceutically acceptable salt thereof.
 58. The method of claim 45, wherein R¹ is methyl; R² is hydrogen; R⁴ and R⁵ are CF₃; A is

and R³ is selected from

or a pharmaceutically acceptable salt thereof.
 59. The method of claim 45, wherein R¹ is methyl; R² is hydrogen; R⁴ is CF₃; A is

and R³ is selected from

or a pharmaceutically acceptable salt thereof.
 60. The method of claim 45, wherein R¹ is methyl or cyclopropyl; R² is hydrogen; R⁴ and R⁵ are CF₃; A is

and R³ is selected from

or a pharmaceutically acceptable salt thereof.
 61. The method of claim 45, wherein the compound is selected from the group consisting of: 4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 6-[4-[(4-Fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-[(5-Fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide; 6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 6-[4-Cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 6-[4-(1,2-Dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-(1-methylcyclopropyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-1H-pyridin-2-one; and N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)benzenesulfonamide, or a pharmaceutically acceptable salt thereof. 